Menstrual blood-derived mesenchymal stem cells combined with collagen I gel as a regenerative therapeutic strategy for degenerated disc after discectomy in rats.

BACKGROUND: Annulus fibrosis (AF) defects have been identified as the primary cause of disc herniation relapse and subsequent disc degeneration following discectomy. Stem cell-based tissue engineering offers a promising approach for structural repair. Menstrual blood-derived mesenchymal stem cells (MenSCs), a type of adult stem cell, have gained attention as an appealing source for clinical applications due to their potential for structure regeneration, with ease of acquisition and regardless of ethical issues. METHODS: The differential potential of MenSCs cocultured with AF cells was examined by the expression of collagen I, SCX, and CD146 using immunofluorescence. Western blot and ELISA were used to examine the expression of TGF-ß and IGF-I in coculture system. An AF defect animal model was established in tail disc of Sprague-Dawley rats (males, 8 weeks old). An injectable gel containing MenSCs (about 1*106/ml) was fabricated and transplanted into the AF defects immediately after the animal model establishment, to evaluate its repairment properties. Disc degeneration was assessed via magnetic resonance (MR) imaging and histological staining. Immunohistochemical analysis was performed to assess the expression of aggrecan, MMP13, TGF-ß and IGF-I in discs with different treatments. Apoptosis in the discs was evaluated using TUNEL, caspase3, and caspase 8 immunofluorescence staining. RESULTS: Coculturing MenSCs with AF cells demonstrated ability to express collagen I and biomarkers of AF cells. Moreover, the coculture system presented upregulation of the growth factors TGF-ß and IGF-I. After 12 weeks, discs treated with MenSCs gel exhibited significantly lower Pffirrmann scores (2.29 ± 0.18), compared to discs treated with MenSCs (3.43 ± 0.37, p < 0.05) or gel (3.71 ± 0.29, p < 0.01) alone. There is significant higher MR index in disc treated with MenSCs gel than that treated with MenSCs (0.51 ± 0.05 vs. 0.24 ± 0.04, p < 0.01) or gel (0.51 ± 0.05 vs. 0.26 ± 0.06, p < 0.01) alone. Additionally, MenSCs gel demonstrated preservation of the structure of degenerated discs, as indicated by histological scoring (5.43 ± 0.43 vs. 9.71 ± 1.04 in MenSCs group and 10.86 ± 0.63 in gel group, both p < 0.01), increased aggrecan expression, and decreased MMP13 expression in vivo. Furthermore, the percentage of TUNEL and caspase 3-positive cells in the disc treated with MenSCs Gel was significantly lower than those treated with gel alone and MenSCs alone. The expression of TGF-ß and IGF-I was higher in discs treated with MenSCs gel or MenSCs alone than in those treated with gel alone. CONCLUSION: MenSCs embedded in collagen I gel has the potential to preserve the disc structure and prevent disc degeneration after discectomy, which was probably attributed to the paracrine of growth factors of MenSCs.

The Functional Study of Response Regulator ArlR Mutants in Staphylococcus Aureus.

Staphylococcus aureus is a major cause of hospital-associated infections worldwide. The organism's ability to form biofilms has led to resistance against current treatment options such as beta-lactams, glycopeptides, and daptomycin. The ArlRS two-component system is a crucial regulatory system necessary for S. aureus autolysis, biofilm formation, capsule synthesis, and virulence. This study aims to investigate the role of the arlR deletion mutant in the detection and activation of S. aureus. We created an arlR deleted mutant and complementary strains and characterized their impact on the strains using partial growth measurement. The quantitative real-time PCR was performed to determine the expression of icaA, and the microscopic images of adherent cells were captured at the optical density of 600 to determine the primary bacterial adhesion. The biofilm formation assay was utilized to investigate the number of adherent cells using crystal violet staining. Eventually, the Triton X-100 autolysis assay was used to determine the influence of arlR on the cell autolytic activities. Our findings indicate that the deletion of arlR reduced the transcriptional expression of icaA but not icaR in the ica operon, leading to decrease in polysaccharide intercellular adhesin (PIA) synthesis. Compared to the wild-type and the complementary mutants, the arlR mutant exhibited decreased in biofilm production but increased autolysis. It concluded that the S. aureus response regulatory ArlR influences biofilm formation, agglutination, and autolysis. This work has significantly expanded our knowledge of the ArlRS two-component regulatory system and could aid in the development of novel antimicrobial strategies against S. aureus.

A study of long-term toxicity of multiple mixtures with hormetic effects by the characteristic parameter σ2(k∙ECx) and stepwise method.

A previous study found that the characteristic parameter σ2(k∙ECx) (the concentration ECx and slope k of the concentrationresponse curve (CRC) at the effect x %) can predict the acute combined toxicity of multiple mixtures with S-shaped CRCs. In this paper, the competence of σ2(k∙ECx) to predict the long-term toxicity of multiple mixtures with J-shaped CRCs was explored using the Aliivibrio fischeri as the test organism. The combined toxicity was evaluated by the independent action (IA) model and the effect ratio (ERx) model. The stepwise method was used to divide J-shaped CRC into ML and MR (SL and SR). The results showed that the σ2(k∙ECx) and ERx of each segment was in good agreement with the exponential function. A new type of mixture was added to the original type A and type B, whose rules of interaction were opposite to those of type B (named opposite B, OB). This paper improves the understanding and analysis of the J-shaped CRCs in environmental risk assessment.

Joint action of binary mixtures based on parameter k·ECx from concentration-response curves in long-term toxicity assay.

A previous acute toxicity study of binary mixtures showed that the combined toxicity can be predicted with the parameter k∙ECx. To systematically investigate the ability of k∙ECx to predict the chronic combined toxicity of binary mixtures, the toxicity of six contaminants and five binary mixtures was determined by long-term microplate toxicity analysis (L-MTA) using Aliivibrio fischeri as the test organism. The independent action model (IA) and the relative model deviation ratio (rMDR) were employed to determine the relationship between the Δ(k∙ECx)% and rMDRx. The results showed that these two factors conformed to the exponential function in long-term toxicity. Owing to the time-dependence of toxicity, the mixture type of chronic toxicity changes to the relative type of acute toxicity. If the acute toxicity of binary mixtures changes their mode of joint action throughout the concentration range, the chronic toxicity will also change their mode of joint action, and vice versa. This study clarified the change rules of the joint action of binary mixtures in acute and chronic toxicity which can promote research on chronic toxicity of binary mixtures.

Study of the joint action of multi-component mixtures based on parameter σ2(k∙ECx) characterizing the shape difference of concentration-response curves.

A previous study has revealed that the parameter k∙ECx, characterizing the shape of concentration-response curves (CRCs), could predict the combined toxicity of binary mixtures. This study further explored the predictability of multi-component mixtures. Eleven component mixtures were designed using the uniform design ray, and the acute toxicity of the eleven environmental pollutants and their mixtures to Vibrio fischeri was determined using microplate toxicity analysis. We used independent action (IA) and the effect residual ratio (ERRx) models to evaluate the combined toxicity of multi-component mixtures and ascertain the functional relationship between σ2(k∙ECx), a parameter characterizing the CRC morphological difference of multi-component mixtures, and combined toxicity. The variance σ2(k∙ECx) of each component characteristic parameter of multi-component mixtures gradually increased in the concentration range, and the relationship between σ2(k∙ECx) and ERRx was consistent with the exponential function. The literature verification showed that this rule is generally applicable to the acute toxicity of multi-component mixtures to luminescent bacteria. The exponential function showed the variation rule of the joint action of multi-component mixtures. In the present study, the joint toxicity of multi-component mixtures can be predicted from single toxicity and small amount of multiple toxicity, circumventing complex multi-component toxicity experiments.

Enhancements of short-chain fatty acids production via anaerobic fermentation of waste activated sludge by the combined use of persulfate and micron-sized magnetite.

The combination of persulfate (PDS) with micron-sized magnetite (Fe3O4) was introduced into the process of anaerobic fermentation (AF) to scrutinize the short chain fatty acid (SCFA) production from waste activated sludge for the first time. The synergetic effect of PDS and Fe3O4 results in the promotion of intracellular and extracellular substance liberation, augment in key hydrolases activities, and enrichment of hydrolytic and acidifying microbial population. Meanwhile, carbohydrate, amino acid, and energy metabolism as well as enzymes, are considerably accelerated. Consequently, the maximum SCFAs yield is significantly enhanced to 391.25 mg COD/L on day 8 of AF with the addition of 0.3 g Fe3O4/g SS and 0.5 g PDS/g SS, which was 2.39-folds than that of the control. The relative abundance of Actinobacteria were highly enriched and reached to 35.76% at the class level. This work affords an effective avenue to evidently boost the production of SCFAs from WAS via AF.

Development of genic KASP SNP markers from RNA-Seq data for map-based cloning and marker-assisted selection in maize.

BACKGROUND: Maize is one of the most important field crops in the world. Most of the key agronomic traits, including yield traits and plant architecture traits, are quantitative. Fine mapping of genes/ quantitative trait loci (QTL) influencing a key trait is essential for marker-assisted selection (MAS) in maize breeding. However, the SNP markers with high density and high polymorphism are lacking, especially kompetitive allele specific PCR (KASP) SNP markers that can be used for automatic genotyping. To date, a large volume of sequencing data has been produced by the next generation sequencing technology, which provides a good pool of SNP loci for development of SNP markers. In this study, we carried out a multi-step screening method to identify kompetitive allele specific PCR (KASP) SNP markers based on the RNA-Seq data sets of 368 maize inbred lines. RESULTS: A total of 2,948,985 SNPs were identified in the high-throughput RNA-Seq data sets with the average density of 1.4 SNP/kb. Of these, 71,311 KASP SNP markers (the average density of 34 KASP SNP/Mb) were developed based on the strict criteria: unique genomic region, bi-allelic, polymorphism information content (PIC) value ≥0.4, and conserved primer sequences, and were mapped on 16,161 genes. These 16,161 genes were annotated to 52 gene ontology (GO) terms, including most of primary and secondary metabolic pathways. Subsequently, the 50 KASP SNP markers with the PIC values ranging from 0.14 to 0.5 in 368 RNA-Seq data sets and with polymorphism between the maize inbred lines 1212 and B73 in in silico analysis were selected to experimentally validate the accuracy and polymorphism of SNPs, resulted in 46 SNPs (92.00%) showed polymorphism between the maize inbred lines 1212 and B73. Moreover, these 46 polymorphic SNPs were utilized to genotype the other 20 maize inbred lines, with all 46 SNPs showing polymorphism in the 20 maize inbred lines, and the PIC value of each SNP was 0.11 to 0.50 with an average of 0.35. The results suggested that the KASP SNP markers developed in this study were accurate and polymorphic. CONCLUSIONS: These high-density polymorphic KASP SNP markers will be a valuable resource for map-based cloning of QTL/genes and marker-assisted selection in maize. Furthermore, the method used to develop SNP markers in maize can also be applied in other species.

Prediction of the joint action of binary mixtures based on characteristic parameter k∙ECx from concentration-response curves.

The evaluation of joint toxicity of mixtures is an important topic in toxicology. Previous studies have found that the parameter k∙ECx of concentration response curves (CRCs) can be used to assess the applicability of concentration addition model (CA). This study further assesses the predictability of k∙ECx on the joint toxicity evaluation. The toxicities of the twelve environmental pollutants, as well as those of binary mixtures with an equivalent-effect concentration ratio, to Vibrio fischeri were determined by using the microplate toxicity analysis. The toxicity evaluation of mixtures was conducted by CA and independent action model (IA). The relationship between the joint toxicity (measured by the relative model deviation ratio (rMDR)) and the k∙ECx was studied. The results shows that the k∙ECx could reflect the shape of CRCs in the whole concentration range. According to the IA and CA, 65% of the mixtures produce strong antagonistic or synergistic effect due to the significant difference of k∙ECx. The percentage of the relative difference of k∙ECx of components and the rMDRx can be fitted by an exponential function. Different types of interactions could be described using this function. It is suggested that the joint toxicity of binary mixtures can be assessed with the parameter k∙ECx, which can quickly get very important data when planning experiments, but also reduce the number of experiments.

Development and Characterization of Nanobodies Targeting the Kupffer Cell.

Nanobodies that are derived from single-chain antibodies of camelids have served as powerful tools in diagnostics, therapeutics and investigation of membrane receptors' structure and function. In this study, we developed a series of nanobodies by a phage display screening building from lymphocytes isolated from an alpaca immunized with recombinant mouse Kupffer cell receptor Clec4F, which is involved in pathogen recognition by binding to galactose and N-acetylgalactosamine. Bio-panning selections retrieved 14 different nanobodies against Clec4F with an affinity ranging from 0.2 to 2 nM as determined by SPR. Those nanobodies mainly recognize 4 different epitopes as analyzed via competitive epitope binning. By analysis of the radioactivity in each organ after injection of 99mTc labeled Clec4F nanobodies in naïve mice, we found that these nanobodies are targeting the liver. Furthermore, we performed a structural characterization at atomic resolution of two of the Clec4F nanobodies from different epitope groups, which revealed distinct features within the CDR2 and CDR3 regions. Taken together, we developed a series of nanobodies targeting multiple distinct recognition epitopes of the Kupffer cell-specific receptor Clec4F which may be useful for its structural and functional investigation as well as for use as molecular imaging and therapeutic agents.

Fe2O3 enhanced high-temperature arsenic resistance of CeO2-La2O3/TiO2 catalyst for selective catalytic reduction of NO x with NH3.

High-temperature arsenic resistance catalysts of CeLa0.5Fe x /Ti (x = 0, 0.1, 0.2, 0.3, 0.4, 0.5) series were prepared and measured under a simulation condition of arsenic poisoning. The as-prepared catalysts were characterized by XRD, SEM, TEM, and XPS. The specific surface area and pore size of the catalysts were measured. At x = 0.2, the catalyst shows the best arsenic resistance and catalytic performance. The active temperature range of the CeLa0.5Fe0.2/Ti catalyst is 345-520 °C when the gas hourly space velocity is up to 225 000 mL g-1 h-1. Compared with commercial vanadium-based catalysts, CeLa0.5Fe0.2/Ti shows much better catalytic performance. The introduction of Fe will improve the dispersion of CeO2 and increase the concentration of Ce3+ and unsaturated active oxygen on the surface. The NH3-TPD and H2-TPR results show that the CeLa0.5Fe0.2/Ti catalyst has more acidic sites and more excellent redox performance than CeLa0.5Fe0/Ti. The CeLa0.5Fe0.2/Ti catalyst might have application prospects in the field of selective catalytic reduction of NO x with NH3.

Biomimetic Metal-Organic Framework Composite-Mediated Cascade Catalysis for Synergistic Bacteria Killing.

Encapsulating nanoparticles/biomolecules into metal-organic freamworks (MOFs) has proven to be highly effective in creating new functions during their applications. However, it is highly desirable yet remains challenging to achieve the synergy of specific functions between the MOF host and guest species. Herein, inspired by the natural multienzyme system, a novel MOF composite biomimetic structure based on the coencapsulation of glucose oxidase (GOx) and l-arginine (l-Arg) into Cu-MOFs (CuBDC) with Fenton-like catalytic activity is designed for achieving the synergistic antibacterial effect. Once activated by GOx-catalyzed glucose oxidation, a large amount of oxygen radicals, toxic ONOO-, and NO are rapidly produced over this well-designed l-Arg/GOx@CuBDC through a double-cascade reaction. Thanks to the synergy of highly reactive species, outstanding antibacterial effects (bacterial inactivation ≥97%) are observed at very low doses (38 µg mL-1 for Escherichia coli and 3.8 µg mL-1 for Staphylococcus aureus). In addition, the in vivo experiment in mice demonstrated that the as-prepared l-Arg/GOx@CuBDC has good biocompatibility, indicating its good potential in practical applications. Such a biomimetic multienzyme system proposes a new design idea for highly efficient antibiosis as well as even therapy for tumors.

Low Level of Vitamin D is a Risk Factor for the Occurrence of Early and Late Onset Pre-Eclampsia in Pregnant Women.

BACKGROUND: This study was aimed to investigate the relationship between 25-hydroxy vitamin D (25(OH)D) level and the occurrence of pre-eclampsia (PE) and also the risk factors of developing early and late onset PE. METHODS: A total of 370 pregnant women were included between January 2015 and December 2016 at our hospital. PE was defined as the presence of maternal blood pressure > 140/90 mmHg and 24-hour proteinuria levels > 300 mg or 2 + in a random sample of urine after the 20th week of pregnancy. Controls were pregnant women without hypertension and proteinuria. Assessment of 25(OH)D was performed at 16 - 20 weeks of gestation. Univariate and multivariate analyses were used to evaluate the association of vitamin D with PE. RESULTS: There were 201 patients with PE while 169 pregnant women were controls. Patients with PE had older maternal age (p < 0.001), earlier gestation age (p < 0.001), and higher systolic blood pressure (SBP) and diastolic blood pressure (DBP) (p < 0.001). The level of 25(OH)D in the PE group (17.26 ± 13.95 µg/L) was significantly lower than that in controls (22.15 ± 12.65 µg/L, p = 0.019). Moreover, the proportion of 25(OH)D deficiency in patients with PE was significantly higher than that of controls (27.6% vs. 0.9%, p < 0.001). Older age, high SBP, and low level of 25(OH)D were independent risk factors of both early and late onset PE during pregnancy. CONCLUSIONS: Low 25(OH)D level was more likely presented in PE patients and was an independent risk factor of both early and late onset PE.

Spectroscopy, Thermodynamics and Molecular Docking of Fraxinellone with DNA.

Fraxinellone is an important botanical lactone compound and has been demonstrated to have insecticidal activity. To provide theoretical support to the assessment on the safety of utilizing fraxinellone as a natural insecticidal agent, the interactions between fraxinellone and armyworm DNA, salmon sperm DNA and calf thymus DNA were investigated using UV-Vis absorption spectroscopy, isothermal titration calorimetry, and molecular docking. Results showed that there were two types of combinations between fraxinellone and three kinds of DNA. Type I combination had an equilibrium constant of combination (Ka1) of about 105 and binding sites (n1) of 0.40-0.70, while type II combination had an equilibrium constant of combination (Ka2) of 103 and binding sites (n2) of 1.35-3.15. Results of molecular docking showed that there were non-classical embedding type interactions between fraxinellone and three kinds of DNA, with the reaction taking place in small groove areas of the DNA structure, resulting in relatively weak interactive forces.

Novel foetal echocardiographic image processing software (5D Heart) improves the display of key diagnostic elements in foetal echocardiography.

BACKGROUND: To evaluate the clinical value of foetal intelligent navigation echocardiography (5D Heart) for the display of key diagnostic elements in basic sections. METHODS: 3D volume datasets of 182 normal singleton foetuses were acquired with a four chamber view by using a volume probe. After processing the datasets by using 5D Heart, eight cardiac diagnostic planes were demonstrated, and the image qualities of the key diagnostic elements were graded by 3 doctors with different experiences in performing foetal echocardiography. RESULTS: A total of 231 volume datasets acquired from the 182 normal foetuses were used for 5D Heart analysis and display. The success rate of 8 standard diagnostic views was 88.2%, and the success rate of each diagnostic view was 55.8-99.2% and 70.7-99.0% for the random four chamber view as the initial section and for the apical four chamber view as the initial section, respectively. The success rate of each diagnostic element in the 8 diagnostic sections obtained by 5D Heart was 58.9%~ 100%. Excellent agreement was found between experienced sonographers and less-experienced sonographers (kappa> 0.769). Inter- and intra-observer agreement were substantial to near-perfect, kappa values ranging from 0.612 to 1.000 (Cohen's kappa). CONCLUSIONS: 5D Heart can significantly improve the image quality of key diagnostic elements in foetal echocardiography with low operator dependency and good reproducibility.

Effect of soil microorganisms and labile C availability on soil respiration in response to litter inputs in forest ecosystems: A meta-analysis.

Litter inputs can influence soil respiration directly through labile C availability and, indirectly, through the activity of soil microorganisms and modifications in soil microclimate; however, their relative contributions and the magnitude of any effect remain poorly understood. We synthesized 66 recently published papers on forest ecosystems using a meta-analysis approach to investigate the effect of litter inputs on soil respiration and the underlying mechanisms involved. Our results showed that litter inputs had a strong positive impact on soil respiration, labile C availability, and the abundance of soil microorganisms, with less of an impact related to soil moisture and temperature. Overall, soil respiration was increased by 36% and 55%, respectively, in response to natural and doubled litter inputs. The increase in soil respiration induced by litter inputs showed a tendency for coniferous forests (50.7%)> broad-leaved forests (41.3%)> mixed forests (31.9%). This stimulation effect also depended on stand age with 30- to 100-year-old forests (53.3%) and ≥100-year-old forests (50.2%) both 1.5 times larger than ≤30-year-old forests (34.5%). Soil microbial biomass carbon and soil dissolved organic carbon increased by 21.0%-33.6% and 60.3%-87.7%, respectively, in response to natural and doubled litter inputs, while soil respiration increased linearly with corresponding increases in soil microbial biomass carbon and soil dissolved organic carbon. Natural and doubled litter inputs increased the total phospholipid fatty acid (PLFA) content by 6.6% and 19.7%, respectively, but decreased the fungal/bacterial PLFA ratio by 26.9% and 18.7%, respectively. Soil respiration also increased linearly with increases in total PLFA and decreased linearly with decreases in the fungal/bacterial PLFA ratio. The contribution of litter inputs to an increase in soil respiration showed a trend of total PLFA > fungal/bacterial PLFA ratio > soil dissolved organic carbon > soil microbial biomass carbon. Therefore, in addition to forest type and stand age, labile C availability and soil microorganisms are also important factors that influence soil respiration in response to litter inputs, with soil microorganisms being more important than labile C availability.

Trimeric structure of the mouse Kupffer cell C-type lectin receptor Clec4f.

The C-type lectin receptor Clec4f has been identified as a specific surface marker for Kupffer cells, although its ortholog is absent in humans and its biological function remains elusive. Here, we report the crystal structure of a truncated mouse trimeric Clec4f. The orientation between the carbohydrate-recognition domain of Clec4f and its neck region differs from other C-type lectins, resulting in an observed distance of 45 Å between the glycan-binding sites within the Clec4f trimer. Interestingly, the trimeric coiled-coil interface within its heptad neck region contains multiple polyglutamine interactions instead of the predominantly hydrophobic leucine zipper found in other C-type lectin receptors. The Clec4f trimeric structure displays unique features regarding its assembly and ligand recognition, shedding light on the evolution and diversity of the C-type lectin family.

Optimization of biochar preparation from the stem of Eichhornia crassipes using response surface methodology on adsorption of Cd2.

In this study, preparation of Eichhornia crassipes stem biochar (ECSBC) was optimized and applied for the removal of Cd2+ from aqueous solution. To obtain the best adsorption capacity of ECSBC, the response surface methodology (RSM) was used to optimize the preparation conditions of ECSBC (OECSBC). The interactions among heating time (X1), heating temperature (X2) and heating rate (X3) were designed by Box-Behnken Design (BBD) experiments. The software gave seventeen runs experiment within the optimal conditions towards two response variables (removal rate and adsorption capacity for Cd2+). The results showed that the mathematical model could fit the experimental data very well and the significance of the influence factors followed the order as heating temperature (X2) > heating rate (X3) > heating time (X1), and the influence of interaction term is: X1 and X2 (heating time and heating temperature) > X2 and X3 (heating temperature and heating rate) > X1 and X3 (heating time and heating rate). Based on the analysis of variance and the method of numerical expected function, the optimal conditions were heating time of 2.42 h, heating temperature of 393 °C, and heating rate of 15.56 °C/min. Under the optimum conditions, the predicted the maximum removal rate and adsorption capacity were 85.2724% and 21.168 mg/g, respectively, and the experimental value of removal rate and adsorption capacity for Cd2+ were 80.70% and 20.175 mg/g, respectively, the deviation from the predicted value were 5.36% and 4.69%. The results confirmed that the RSM can optimize the preparation conditions of ECSBC, and the adsorption capacity of OECSB was improved.

Deciphering the activation and recognition mechanisms of Staphylococcus aureus response regulator ArlR.

Staphylococcus aureus ArlRS is a key two-component regulatory system necessary for adhesion, biofilm formation, and virulence. The response regulator ArlR consists of a C-terminal DNA-binding effector domain and an N-terminal receiver domain that is phosphorylated by ArlS, the cognate transmembrane sensor histidine kinase. We demonstrate that the receiver domain of ArlR adopts the canonical α5ß5 response regulator assembly, which dimerizes upon activation, using beryllium trifluoride as an aspartate phosphorylation mimic. Activated ArlR recognizes a 20-bp imperfect inverted repeat sequence in the ica operon, which is involved in intercellular adhesion polysaccharide production. Crystal structures of the inactive and activated forms reveal that activation induces a significant conformational change in the ß4-α4 and ß5-α5-connecting loops, in which the α4 and α5 helices constitute the homodimerization interface. Crystal structures of the DNA-binding ArlR effector domain indicate that it is able to dimerize via a non-canonical ß1-ß2 hairpin domain swapping, raising the possibility of a new mechanism for signal transduction from the receiver domain to effector domain. Taken together, the current study provides structural insights into the activation of ArlR and its recognition, adding to the diversity of response regulation mechanisms that may inspire novel antimicrobial strategies specifically targeting Staphylococcus.

NIRF-Molecular Imaging with Synovial Macrophages-Targeting Vsig4 Nanobody for Disease Monitoring in a Mouse Model of Arthritis.

Nanobody against V-set and Ig domain-containing 4 (Vsig4) on tissue macrophages, such as synovial macrophages, could visualize joint inflammation in multiple experimental arthritis models via single-photon emission computed tomography imaging. Here, we further addressed the specificity and assessed the potential for arthritis monitoring using near-infrared fluorescence (NIRF) Cy7-labeled Vsig4 nanobody (Cy7-Nb119). In vivo NIRF-imaging of collagen-induced arthritis (CIA) was performed using Cy7-Nb119. Signals obtained with Cy7-Nb119 or isotope control Cy7-NbBCII10 were compared in joints of naive mice versus CIA mice. In addition, pathological microscopy and fluorescence microscopy were used to validate the arthritis development in CIA. Cy7-Nb119 accumulated in inflamed joints of CIA mice, but not the naive mice. Development of symptoms in CIA was reflected in increased joint accumulation of Cy7-Nb119, which correlated with the conventional measurements of disease. Vsig4 is co-expressed with F4/80, indicating targeting of the increasing number of synovial macrophages associated with the severity of inflammation by the Vsig4 nanobody. NIRF imaging with Cy7-Nb119 allows specific assessment of inflammation in experimental arthritis and provides complementary information to clinical scoring for quantitative, non-invasive and economical monitoring of the pathological process. Nanobody labelled with fluorescence can also be used for ex vivo validation experiments using flow cytometry and fluorescence microscopy.

A nano-reactor based on PtNi@metal-organic framework composites loaded with polyoxometalates for hydrogenation-esterification tandem reactions.

Tandem catalysis (i.e., a process in which a desirable product is synthesized by a one-step process consisting of sequential reactions) has attracted intensive attention owing to its sustainable green and atom-economical characteristics. In this process, the utilization of a high-efficiency multifunctional catalyst is key. However, different functional sites integrated within the catalyst are required to be rationally designed and precisely engineered to guarantee the synergy between the catalytic reactions. Herein, a novel kind of hydrogenation-esterification tandem catalyst with metal/acid (alloy/polyoxometalates) active sites integrated within the metal-organic frameworks (MOFs) was prepared by a facile self-sacrificial template route. In this tandem catalyst, the MOF cavities served as tandem reactors, the PtNi alloy sites encapsulated within the MOF material acted as hydrogenation sites, and the solid phosphotungstic acid embedded in the MOF cavities provided esterification sites. This well-designed tandem catalyst showed outstanding activity and selectivity towards the one-step synthesis of amino-ester-type anesthetics (e.g., benzocaine) owing to the synergistic catalysis of the metal and acid sites. Clearly, this novel tandem catalyst simplifies the traditional industry process and provides a new method to rationally construct new tandem catalysts.